Prohibitin: A Novel Molecular Player in KDEL Receptor Signalling

Figure 5

Trafficking of VSVG in PHB knock-down cells. (a) Depletion of PHB does not interfere with transport of VSVG to the Golgi complex, but it impairs VSVG arrival at the plasma membrane. HeLa-myc cells were treated with scrambled siRNAs (Control) or with siRNAs against PHB (PHB siRNA,) for 96 h. After infection with VSV for 45 min, the cells were incubated at 40°C for 3 h (temperature block) and then shifted to 32°C for the indicated times (temperature-block release). Left panels: The cells were fixed and stained for VSVG (green) and GM130 (marker for Golgi area definition, red); the merged images are also shown (Total VSVG/GM130). Right panels: Immunostaining for total VSVG (green), VSVG at the plasma membrane (revealed by an antibody against the extracellular domain of VSVG; External VSVG, red). The merged images shown include immunostaining for GM130 (blue). Scale bars, 10 μm. (b) Quantification of VSVG localised at the plasma membrane of HeLa-myc cells treated as in (a). Data are means ± SEM of VSVG immunofluorescence on the plasma membrane, from three independent experiments, with at least 50 cells quantified per experiment. compared to control cells (-test).

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